www.nature.com/scientificreports OPEN A biomolecular anthropological investigation of William Adams, the frst SAMURAI from England Fuzuki Mizuno1*, Koji Ishiya2,3, Masami Matsushita4, Takayuki Matsushita4, Katherine Hampson5, Michiko Hayashi1, Fuyuki Tokanai6, Kunihiko Kurosaki1* & Shintaroh Ueda1,5 William Adams (Miura Anjin) was an English navigator who sailed with a Dutch trading feet to the far East and landed in Japan in 1600. He became a vassal under the Shogun, Tokugawa Ieyasu, was bestowed with a title, lands and swords, and became the frst SAMURAI from England. "Miura" comes from the name of the territory given to him and "Anjin" means "pilot". He lived out the rest of his life in Japan and died in Hirado, Nagasaki Prefecture, in 1620, where he was reportedly laid to rest. Shortly after his death, graveyards designated for foreigners were destroyed during a period of Christian repression, but Miura Anjin’s bones were supposedly taken, protected, and reburied. Archaeological investigations in 1931 uncovered human skeletal remains and it was proposed that they were those of Miura Anjin. However, this could not be confrmed from the evidence at the time and the remains were reburied. In 2017, excavations found skeletal remains matching the description of those reinterred in 1931. We analyzed these remains from various aspects, including genetic background, dietary habits, and burial style, utilizing modern scientifc techniques to investigate whether they do indeed belong to the frst English SAMURAI. William Adams was born in Gillingham, England in 1564. He worked in a shipyard from the age of 12, and later studied shipbuilding techniques, astronomy and navigation. He also experienced Arctic expeditions and naval captaincy. At the age of 34, he embarked on an expedition to the orient by a private Dutch feet as chief ofcer. Te voyage encountered numerous disasters, and of the fve ships that set sail, only the one in which he eventually sailed on, the Liefde, remained. With a few survivors onboard the Liefde landed in present-day Oita Prefecture in Japan in 1600. Te Shogun, Tokugawa Ieyasu seized power and established a government at Edo (now known as Tokyo) in 1603. Ieyasu took on Adams as an advisor and studied mathematics and geography from him. He also heavily relied on Adams as a diplomatic advisor to the Shogunate. Adams was requested by Ieyasu to build a Western-style sailing ship, and in 1604 he built the frst shipbuilding dock in Japan in Ito. In 1607, in response to Adams’s achievements, Ieyasu selected him for the high-prestige position as a direct retainer in the Shogun’s court, entrusting him with Miura-gun (now a part of Yokosuka City) territories and swords. He was bestowed the name Miura Anjin. "Miura" comes from the name of the territory, and "Anjin" means "pilot". Tis made him the frst Englishman to hold the position of SAMURAI. He had the unusual situation of living as a Japanese SAMU- RAI despite being a foreigner. He worked for the Shogunate’s overseas trade industry, including the establishment of the Hirado Trading House in the Netherlands and the United Kingdom (1613). When the British East India Company ship Clove arrived in Hirado in 1614 Adams had a chance to return to England, but he opted to stay in Japan to live out the rest of his days. Adams reportedly died in 1620 in Hirado, Nagasaki Prefecture (Fig. 1a), and was buried in a cemetery designated for foreigners in 1620, due to the changing times in Japan in the Early Edo period (1600–1868). Afer the Shimabara rebellion in 1637, shortly afer Adams’ death, foreign graveyards were destroyed during a period of Christian repression. Legend says that his bones were, however, saved by a close family friend and reburied. In 1931, a grave marked as a Miura family tomb was excavated1. During the 1Department of Legal Medicine, Toho University School of Medicine, 5-21-16, Omori-Nishi, Ota-ku, Tokyo 143-8540, Japan. 2Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Sapporo 062-8517, Japan. 3Computational Bio Big Data Open Innovation Lab (CBBD-OIL), National Institute of Advanced Industrial Science and Technology (AIST)-Waseda University, Tokyo 169-8555, Japan. 4The Organization of Anthropological Research, Yamaguchi 759-6604, Japan. 5Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan. 6Center for Accelerator Mass Spectrometry, Yamagata University, Kaminoyama 999-3101, Japan. *email: fuzuki.mizuno@ med.toho-u.ac.jp; [email protected] Scientifc Reports | (2020) 10:21651 | https://doi.org/10.1038/s41598-020-78723-2 1 Vol.:(0123456789) www.nature.com/scientificreports/ Figure 1. (a) A map showing the location of Hirado City, Nagasaki Prefecture, Japan, where Miura Anjin’s grave is situated. Base map sourced from https ://www.freem ap.jp. (b) An image of the tombstone at Miura Anjin’s grave. excavation, fragments of crania, scapulae, vertebrae, ribs, femurs, and one molar were unearthed along with one piece of earthenware, and 30 corroded nails. Te bones were found directly under the gravestone in order; crania–upper body–upper limbs–spine–pelvis–lower limbs, and it was estimated that they were interred in a cofn (around 190 cm long and 65 cm wide). From the position of the head and feet, it was recorded that the individual would have been around 170 cm tall1. Afer the excavation, the bones were reburied at the current site, where a tombstone dedicated to Miura Anjin (William Adams) now stands (Fig. 1b). During archaeological excavations in 2017, human skeletal remains matching the description of those found in 1931 were uncovered. Results and discussion Approval for the present study was provided by the Ethics Committee of Toho University School of Medicine (A18101_A18058) and relevant guidelines were followed for the study. Site and excavation. Archaeological excavations were conducted in Hirado City, Nagasaki Prefecture, Kyushu, Japan in 2017 (Fig. 1a). During excavations directly under the tombstone (Fig. 1b), a porcelain vase was excavated from under a layer of gravel, and skeletal remains belonging to a single individual were found inside of it. Te skeletal remains consisted of the skull, mandible, femur, and part of the tibia, and match the descrip- tion of those from the 1931 report. Inspection of the layer under the burial revealed markings of a rectangular grave (Fig. S1ab). Te remains consist of a part of the cranium, mandible, femur, tibia, and bone fragments (Fig. 2). Tere were no accompanying grave goods indicating the identity of the remains. However, the grave pit detected in 1931 was a rectangular cuboid shape, and so it is probable that a cofn was used, and the burial posture was extended supine. In the Edo period, circular shape graves were the norm, and extended supine burials were only used for Westerners or Christians, not for most native Japanese. Te grave matches the description of a burial pit with the remains of skeletal remains and a cofn found and reinterred in 1931 1. In addition, the grave site has been protected as one of a nobleman by the people living in Hirado for centuries. Scientifc Reports | (2020) 10:21651 | https://doi.org/10.1038/s41598-020-78723-2 2 Vol:.(1234567890) www.nature.com/scientificreports/ Figure 2. Te fragments of human bone recovered during the excavation are colored blue. Te diagram shows the mandible at the top, the skull in the middle, and the long bones below. Radiocarbon dating and stable isotopic measurements. Te stable carbon isotope ratio was − 18.87‰ and the stable nitrogen isotope ratio was + 11.95‰ for the collagen sample. Te carbon content of the collagen was 15.82%, the nitrogen content was 4.85%, and the atomic carbon/nitrogen ratio was 3.8. Tis value was diferent from the recommended one (2.9–3.6) obtained for well-preserved bone collagen2. Tis result indicates that the sample was afected by contamination of biotic substances such as humic acids. Te radiocarbon age was dated as 410 ± 30 years BP (Beta-495523, Conventional 14C age). Since humans intake some of their proteins from marine products, bone collagen contains some marine carbon. Hence, it is necessary to consider a marine reservoir efect to obtain a calendar age from bone collagen. Te ratio of marine resources was set to 25.1% from the carbon isotope ratio (δ13C) following the methodology of Arneborg et al.3. Te ratio is applied to obtain the modelled calibration curve, which comprises a combination of the international calibration datasets of IntCal204 and Marine205. Ten, the calendar age is calculated using OxCal 4.46. Te infu- ence of the local marine reservoir was taken from the data for nearby Hakata Bay, HKA2-1 1.0–1.2 m7. Te local marine reservoir efect is ΔR = 53 ± 63 (14C year) which is derived by using the 14 CHRONO Marine Reservoir Database (http://calib .org/delta r/). Te obtained calendar dates (calibrated ages with 95.4% probability) are Scientifc Reports | (2020) 10:21651 | https://doi.org/10.1038/s41598-020-78723-2 3 Vol.:(0123456789) www.nature.com/scientificreports/ calAD 1466–1819 (92.0%), 1836–1867 (1.6%), and 1925–1948 (2.0%) (Fig. S2). Although the atomic C/N ratio is outside the recommended values, the calibrated calendar age is consistent with "Adams died in Hirado, on 16 May 1620, at the age of 55". Te ratios of stable carbon and nitrogen isotopes of collagen extracted from human skeletal remains refect the isotopic ratios of the food ingested by the individual, and thus can be used to estimate dietary habits (e.g., dependence on marine products, C3 plants, C4 plants, etc.). Figure S3 shows plots of the carbon and nitrogen stable isotope ratios on an isotope distribution map.